Isolation and characterization of thermophilic bacteria from Omani hot springs and heterologous expression and purification of potential thermostable enzymes.

Microorganisms can thrive in all possible environments including extreme habitats such as northern and southern polar areas, high saline zones and hydrothermal vent systems. Of particular interest for industrial biotechnology are thermophiles, microorganisms which grow and survive at elevated temperatures, since they act as indispensable sources of thermostable enzymes. Thermostable enzymes such as cellulases, amylases, lipases, proteinases and chitinases have many desirable characteristics which make them attractive targets for a wide range of industrial applications. Commercial applications of enzymes include hydrolyzing biopolymers into smaller components, leather preparation, detergents, diagnostics and food processing. Hot springs are considered favorable habitats for thermophilic microorganisms which can produce novel enzymes with unique properties awaiting to be discovered. Thermostable enzymes offer advantages over other enzymes because they remain active at high temperatures, tolerate proteolysis and severe conditions. The Sultanate of Oman has several natural hot springs which have unique biological characteristics making them attractive subjects for research with potential value for industrial biotechnology purposes. No reports about the isolation of thermophilic bacteria from hot spring in Oman has been published yet and little is known about the occurrence and distribution of thermophiles in the Sultanate. The aim of the present proposal is to isolate, identify and characterize thermophilic microbial diversity from several hot springs in Oman by combining various phenotypic (morphology, microscopic and biochemical) and genotypic methods. Furthermore, a number of enzymatic activities (cellulase, lipase, amylase and proteinase) will be screened from samples collected from hot springs. The potential enzymes will be produced in the native bacteria followed by purification ammonium sulphate purification method and their application in different commercial fields will be evaluated. To maximize the yields to commercial levels, the potential enzymes will be recombinantly overexpressed as fusion proteins with GFP in heterologous established protein expression systems (E. coli and/or L. lactis). The levels of protein expression will be assessed by checking fluorescence of the fusion protein under fluorescent microscopy and the possibility of expressing the protein in large scale will be evaluated.